Micro-ecological active bacteria of microbial species

Micro-ecological living bacteria products are made by members of normal strains of the human body or harmless foreign bacteria with the effect of promoting the growth and activity of normal flora, after culture, collection of bacteria, drying into bacteria powder, added to the suitable auxiliary mixtureUsed to prevent and treat related symptoms and diseases caused by microbiome disordersmicro-ecological living bacteria products must be composed of non-pathogenic live bacteria, whether in the production process, product storage and use of the living bacteria state should be stableIt can be made from one, more or several bacteria unit or multi-price combinationAccording to its different ways of use and methods can be prepared into tablets, capsules, granules or dispersants and other dosage formsbasic requirementsthe preparation method and process of micro-ecological live bacteria products should ensure that the finished product contains sufficient number of live bacteria, maintain its stability, and at the same time prevent the pollution of external factorsProduction and inspection facilities, raw materials and accessories, water, utensils, animals, etcshould comply with the relevant requirements of "all cases"manufacturingproduction ofproduction of bacteria shall comply with the relevant provisions of the "Regulations on the Regulations on the Regulations on the Control of The Use of Bacterial Poisons for the Production of Biological Products"1, name and sourcethe selection of production bacteria should come from the body normal flora, or non-toxic harmless to the human body, with the promotion of normal flora growth and activity of foreign bacteriaThe separation process and transmission background of bacteria should be clear, should have stable biological and genetic characteristics, and be able to maintain a stable state of active bacteria, and have been proven safe and effective by laboratory and clinical trials2, the establishment ofspecies of seed batch, the classification of the species, should be in accordance with the latest version of the Bergry's Manual of Systematic System Identification Manual (Bergry's Manual of Systematic And Inperium) and Berger's Bacterial Naming Manual (Bergry's Manual of Determinedative Bacteria) relevant provisions, including morphological, growth metabolic characteristics inspectionThe original or primary seed should also be examined for genetic characteristics and antibiotic sensitivitythethethe third-level seed batch routine examination includes the following 3 items:(1) culture characteristics and dyeing mirror test inoculated the species in suitable culture medium, set anaerobic or anaerobic environment culture, observe their growth, determine the bacteria as aerobic bacteria or anaerobic Sex bacteria, the shape, size, surface, edge, transparency, color and other characteristics of individual colonies grown on agar flat dishes by dashmethod, and the growth characteristics of the strains under different temperature, pH or different concentrations of sodium chloride solution, etctake fresh culture smears of the species to do Gramstain staining, under a microscope to observe the staining reaction, shape, size and arrangement of the bacteria, bacteria with spores should simultaneously observe the shape, size and location of spores (can also be added to spore staining)The results of the inspection should be consistent with the characteristics of the original species(2) Biochemical Reactions Are carried out according to Appendix XI.V"Bacterial Biochemical Reaction Media" to select the appropriate media or other appropriate methods, the results should be in line with the characteristics of the original species(3) Toxicity test Toxicity test is to check the presence of unsafe factors in the species through animal tests to ensure the safety of human use5 mice weighing 18 to 22g, each abdominal cavity injection 0, 3 ml of fresh bacteria (not less than 1, 0 x 109 CFU/0, 3 ml), continuous observation 3 days, mice should be healthy survival, weight gain; Mice through the mouth of the stomach 0, 5 ml of fresh bacteria (not less than 1, 0 x 109CFU/0, 5 ml), 1 time a day, 3 consecutive days, from the first day of gastric continuous observation to the 7th day, the mice should be healthy survival, weight gain, unless otherwise specified, the original seed or the main seed batch shall be subject to the following checks:(1) bacterial metabolites - fatty acid determination according to gas chromatography (Appendix IIIC) or other appropriate methods, should be in accordance with the characteristics of the species(2) Genetic Characteristics Analysis Can be determined by the content of G -Cmol% of bacterial DNA or other appropriate methods, should be in line with the genetic characteristics of the strain(3) Antibiotic Sensitivity Test Using agar diffusion paper method or other appropriate method to test the antibiotic sensitivity of the strain, should conform to the characteristics of the strain (4) stability test bacteria in the suitable medium, after 30 generations of continuous transmission, the 30th generation of culture sending seed verification, all the test results should be consistent with the characteristics of the original bacteria 4, the preservation of the seed batch the original seeds and the main seeds shall be frozen dry and kept below 8 degrees C, and the working seeds shall be kept at the appropriate temperature powder manufacturing should include seed liquid preparation, large tank culture, harvesting bacteria (or spores) and bacteria drying to make bacteria powder If the production of multi-price products, each bacteria should be cultured separately, the preparation of unit-price bacteria powder 1, the production of seed to open the working seed batch, inoculated in suitable media for multi-level seed amplification, should be smeared to do Gramdd dyeing, under the microscope to observe 5 to 10 vision, bacteria dyeing reaction, form should be consistent and in line with the characteristics of the original species The preparation process should prevent pollution, and the number of transmissions of strains should be in accordance with the regulations 2, production of medium using approved media for production 3, culture using liquid culture The seed liquid under appropriate conditions of culture (including anaerobic or aerobic, temperature, time, etc.), the culture process sampling smear for Grameen staining mirror examination, pH detection, spores need to be tested for spore formation rate, should comply with the provisions After the cultivation of sampling for pure bacteria inspection, if found contamination should be discarded the production of multi-price products of the unit price of bacteria powder, should be cultured separately 4, harvested bacteria and made bacteria powder after culture to centrifugal harvesting wet bacteria, mixed with appropriate dispersants, stabilizers Using vacuum freeze-drying method to dry the bacteria, spores can be used heating and drying method, and then crushed, sieve to make powdered bacteria powder 5, the preservation and validity of the bacteria powder shall be determined by the living bacteria stability test 6, bacteria powder verification according to the "bacteria powder verification" item, in line with the provisions before the semi-finished product preparation semi-finished product 1, the preparation the same work seed batch production of up to 2 batches of unit-price bacteria powder can be mixed with the auxiliary material in the approved proportion and evenly mixed into semi-finished products When formulating multi-price products, each unit price powder, accessories according to the proportion of the formula and the preparation procedure mixed evenly, the preparation process should prevent pollution 2, semi-finished product verification according to the "semi-finished product verification" item, should comply with the provisions the finished product 1, dosage form preparation according to the use of the product, the object of use and the drug route and other factors to determine the dosage form The preparation process shall comply with the relevant dosage forms under Appendix I "General Rules of Preparation" 2, batch finished lot numbers should be determined after the preparation of semi-finished products, the preparation date is the production date The same batch of products, should be the same source, quality is one, according to the requirements of sampling inspection, the whole batch of products can be assessed According to the verification results, the semi-finished product slots should be specified, such as more than 24 hours, should be divided into different sub-batches 3, sub-packing, specifications and packaging products shall be in accordance with the relevant provisions of Appendix I "General Rules of Preparation" Packaging shall comply with the relevant provisions of the Biological Products Packaging Regulations Specifications should meet approved specifications verification the quality verification of micro-ecological live bacteria products should include the detection of bacteria powder, semi-finished products and finished products powder 1, the appearance of should be white, gray white or grayish-yellow powder 2, purpose bacteria inspection take a small amount of bacteria powder to add to the appropriate amount, sterilization physiological sodium chloride solution or other suitable dilution, coating in suitable agar flat dish, under appropriate conditions culture, the growth characteristics of culture and staining mirror inspection characteristics should be in line with the characteristics of the production of bacteria 3, the method sift inspection method scored and the results are judged in Appendix 3 of this General Theory If it does not comply with the regulations, it should be abandoned 4, dry weightlessness the content of residual moisture in the bacteria powder will directly affect the survival of live bacteria, the bacteria powder dry weightlessness check It should be measured in accordance with Appendix VII L or instrument method 5, the number of live bacteria to determine the number of live bacteria per gram of bacteria powder Methods can be found in Appendix 2 of this General Theory semi-finished product seisped semi-finished products must do the miscellaneous inspection, according to the drug route to determine the quality control index of the hybrid inspection Methods and results are judged in Appendix 3 of this General Opinion the finished product verification 1, identification test check whether the target bacteria contained in the finished product conforms to the characteristics of the production of the bacteria That is, according to the above-mentioned "seed batch verification" method for growth characteristics, staining mirror inspection and biochemical reaction inspection, should comply with the provisions For multi-price products, the unit-price bacteria characteristics must be checked one by one 2, physical and chemical inspection (1) appearance according to the dosage form, observe the appearance of the product, color tablet appearance should be complete, smooth, white or white, between the bacteria pink spot; (2) Dry weightlessness according to Appendix VII L or instrument method, weight loss should not exceed 5, 0%, spore products should not exceed 7, 0% (3) particle size dispersants and granules should be checked for granularity (4) loading (weight) difference Each dosage form shall be carried out in accordance with the corresponding provisions of Appendix I "General Rules of Preparation" and shall comply with the regulations (5) collapse time limit Capsules, tablets in accordance with appendix V.C, shall comply with the provisions 3, the number of live bacteria to determine according to the general appendix 2 method to determine the number of live bacteria per gram of products, should comply with the provisions Multi-price products should be measured separately the number of active bacteria at each unit price 4, the of the hybrid inspection is to check the contamination of the finished foreign-sourced microorganisms, in order to ensure the safety of human use method and results are judged on the same as the "hybrid examination" item of semi-finished products 5, safety test safety test is a nonspecific toxicity test carried out through animal test, according to the use of the product and human dose to determine the test method (1) intestinal micro-ecological bio-bacterial products inspection called 2g products, added to 8 ml of physiological sodium chloride solution, mixed evenly With 5 mice weighing 18 to 22g, each mouse through the mouth of the stomach 0, 5 ml, 1 time a day, for 3 consecutive days From the first day of gastric irrigation, continuous observation for 7 days, mice should be healthy survival, weight gain, judged qualified If not qualified, you can select another 10 mice retest 1 time, the determination standard is the same (2) vaginal micro-ecological biopsy products check with 24 to 26 g female mice 5, each mouse vagina built into 10 mg products, 1 time a day, for 3 consecutive days Since the first day of administration, continuous observation for 7 days, mice should be healthy survival, weight gain, vaginal local no redness, secretions and other symptoms, judged to be qualified preserved, transported and valid at approved temperatures From the date of production, according to the approved validity period